核转录因子-κB在急性肺损伤小鼠中的动态表达

目的 研究核转录因子-κB(NF-κB)在脂多糖诱发的急性肺损伤小鼠中的动态表达情况.方法 健康纯种昆明鼠25只,随机分为5组,包括正常对照组(腹腔注射等量生理盐水),4组LPS组(腹腔注射LPS).LPS组在腹腔注射脂多糖后的不同时间点(1、3、9、12h)处死小鼠,检测相关指标.通过动脉血氧分压(PaO2)检测肺功能;普通光镜观察HE染色肺组织的病理变化;Western blotting和免疫组化法检测NF-κB p65在蛋白水平上的表达差异.结果 LPS注射后1h,PaO2明显下降并有炎症反应的病理变化;小鼠肺组织中的NF-κB p65呈双峰表达,峰值分别为1h和9h. 结论 NF-κB在脂多糖所致急性肺损伤的炎症反应中发挥着重要作用.     

大黄素对角膜炎大鼠角膜组织NF-κB活化表达的影响及意义

目的：观察大黄素对角膜炎大鼠角膜组织核因子-κB （NF-κB ）活化表达的影响及意义。方法制备角膜炎动物模型。模型制备前30 min,分别于治疗组和炎症组大鼠结膜下注射大黄素及溶剂。裂隙灯显微镜观察大鼠眼部变化;Western blotting检测评价角膜组织NF-κB 活化表达;免疫组织化学法检测角膜组织细胞间黏附分子-1（ICAM-1）蛋白的表达。结果成功制备了角膜炎大鼠模型,大黄素可改善角膜炎大鼠的眼部炎症反应,减少角膜组织炎细胞浸润。治疗组κB抑制蛋白α（IκBα）在脂多糖（LPS ）刺激后各时点的表达均高于同时点炎症组表达（P＜0．01）。LPS刺激可促进ICAM-1在角膜组织的阳性表达,该作用可被大黄素部分抑制。结论大黄素可部分抑制角膜炎大鼠角膜组织中NF-κB活化,有效减轻角膜炎症的损伤。     

银杏苦内酯B对急性肺损伤小鼠肺组织NF-κB表达的影响

目的 研究银杏苦内酯B(BN52021)对脂多糖(LPS)诱发的急性肺损伤小鼠肺组织中核转录因子κB(NF-κB)蛋白质表达的影响.方法 健康昆明小鼠50只,随机分为2组:LPS组和BN52021预处理组.每组又根据不同的观察时间点(0、1、3、9 h和12 h)随机分为5个业组,每组5只小鼠.通过检测肺湿质量/干质量值(W/D)初步判断肺功能,普通光镜观察HE染色肺组织的病理变化,Western blot检测NF-κB p65在蛋白水平上的表达差异,ELISA法测定肺组织匀浆中肿瘤坏死因子-α(TNF-α)及白细胞介素-10(IL-10)浓度.结果 LPS注射后,LPS组W/D显著升高,提示存在肺水肿;BN52021预处理组在不同时间点的W/D均低于LPS组(P<0.05,P<0.01).LPS注射后1 h即有明显炎症反应的病理变化:两组光镜下均显示肺泡出血及中性粒细胞浸润,但BN52021预处理组明显轻于LPS组.LPS组NF-κB p65呈双峰表达,峰值分别为LPS注射后1 h和9 h;BN52021预处理组NF-κB p65呈单峰表达,峰值出现于9 h,除9 h外,各时间点BN52021预处理组NF-κB 065的表达均低于LPS组(P<0.05).与LPS组相比,BN52021预处理组可以明显降低TNF-α的表达(P<0.05),但2组IL-10峰值无统计学意义.结论 BN52021通过抑制NF-κB的早期活化抑制炎症反应,减轻肺组织损伤.     

非特异性角膜炎症反应中PDTC对LPS诱导LFA-1 mRNA表达的影响

目的：探讨淋巴细胞功能相关抗原-1(LFA-1)的mRNA表达与非特异性角膜炎症反应的相关性及核因子NF-κB抑制剂对LFA-1 mRNA表达的阻断作用。方法：建立BALB/C鼠角膜缝线及联合结膜下药物注射的动物模型,通过逆转录聚合酶链式反应（RT-PCR）,对单纯缝线组、缝线联合脂多糖（LPS）组和缝线联合LPS及吡咯啉烷二硫代氨基甲酸盐（PDTC）组术后1、3、7和14 d不同时相点角膜LFA-1 mRNA的转录水平进行检测。结果：术后第1、3、7 d时,角膜缝线联合LPS处理组LFA-1 mRNA的表达明显高于单纯缝线组(P<0.05);14 d时,角膜缝线联合LPS处理组与单纯缝线组比较差异无显著性（P>0.05）。而1和7 d时缝线联合LPS和PDTC组LFA-1mRNA的表达明显高于缝线联合LPS组;3和14 d时缝线联合LPS和PDTCLFA-1 mRNA的表达明显低于缝线联合LPS组(P<0.05)。结论：联合结膜下注射LPS可在术后诱导LFA-1 mRNA的高水平表达,而核因子NF-κB抑制剂PDTC可抑制角膜LFA-1 mRNA的表达,表明PDTC对LFA-1 mRNA表达具有阻断调控作用。     

曲安奈德对核因子-κB、血管内皮生长因子在大鼠视网膜新生血管中表达的影响

目的:探讨曲安奈德对核因子-κB(NF-κB)及血管内皮生长因子(VEGF)在大鼠视网膜新生血管病变中表达的影响。方法:选择7 d SD幼鼠40只,其中30只大鼠建立氧诱导的视网膜新生血管病变模型,10只置于正常环境中。30只大鼠中20只大鼠出氧箱后给予玻璃体腔内注射曲安奈德1μl(0.04 mg),对侧眼注射平衡盐溶液(balanced saltsolution,BSS)作为对照,分为高氧+药物治疗组和高氧+BSS液对照组,每组20只眼;另10只大鼠作为高氧对照组。视网膜组织切片HE染色,免疫组织化学法检测NF-κB及VEGF的表达。结果:高氧+药物治疗组与高氧对照组及高氧+BSS组相比,新生血管内皮细胞核数目明显减少(P0.01),高氧+药物治疗组的NF-κB及VEGF的表达均低于高氧对照组及高氧+BSS组(P0.05)。VEGF表达升高的同时,NF-κB的表达呈升高趋势。结论:曲安奈德明显抑制NF-κB及VEGF的表达,即有抑制新生血管形成的作用,NF-κB与VEGF的表达呈正相关,在新生血管形成过程中起协同作用。     

骨化三醇对脂多糖炎性大鼠神经保护作用的研究

目的:观察骨化三醇对脂多糖(LPS)炎性大鼠脑组织的保护效应及作用机制。方法:健康雄性成年SD大鼠52只,随机分为三组:空白对照组6只、骨化三醇干预组23只[4μg/(kg.d)×14d]、脂肪乳剂(安慰剂)组23只[4ml/(kg.d)×14d],在第14d给药结束后1h,腹腔注射LPS 10mg/kg制造大鼠全身炎症模型,用凝胶电泳迁移率分析(EMSA)方法检测造模后3、6和9h脑组织核因子-κB(NF-κB),用ELISA方法测肿瘤坏死因子α(TN-α)、白细胞介素-10(IL-10),用苏木精-伊红(H-E)染色观察脑组织神经细胞损伤情况。结果:LPS腹腔注射使大鼠脑组织NF-κB活化、TNF-α表达增加,并随时间的延长而增高,伴有散在的大脑皮质浅表区神经元损害。骨化三醇干预能下调各时间点NFκ-B活性,降低TNF-α含量,并促进IL-10表达。骨化三醇干预减轻了神经细胞损伤程度。结论:大鼠腹腔注射LPS诱导的全身炎症状态能引发脑损伤,骨化三醇干预可能通过调节脑内炎性-抗炎因子的平衡而发挥神经保护作用。     

升降散治疗急性肺损伤作用的实验研究

[目的]探讨升降散治疗急性肺损伤的作用机理.[方法]40只雄性SD大鼠,随机分为正常组、模型组、升降散组(剂量为2 g/kg).给药6 d后,舌下静脉注射脂多糖(LPS)复制急性肺损伤模型,取肺组织切片,免疫组化检测;采集图像,分析微血管内皮细胞核的平均灰度.[结果]升降散组与模型组比较,能显著性抑制微血管内皮细胞核中核因子κB(NF-κB)的表达(P＜0.01).[结论]升降散可通过抑制肺微血管内皮细胞NF-κB的表达,抑制炎症反应对肺组织造成的急性损伤,对急性肺损伤起到防治作用.     

黄芪多糖对高血压病患者血清致伤血管内皮细胞TLR4、NF-κB表达的影响

目的 观察黄芪多糖(APS)对TLR-NF-κB信号转导通路的影响,探讨其对高血压病患者血清损伤的血管内皮细胞的保护机制.方法 用10%的高血压痛患者及健康人血清干预人脐静脉内皮细胞(HUVEC-C)24 h,实时荧光定量(PCR)法检测TLR4 mRNA表达.不同浓度的APS干预脂多糖(LPS)诱导的HUVEC-C TLR4表达(24 h)及核转录因子(NF-κB)活化(2 h),PCR法检测TLR4 mRNA及NF-κB mRNA的表达,免疫印迹(Western-blot)技术检测TLR4蛋白及NF-κB蛋白的表达.结果 血清作用24 h后,高血压病组TLR4 mRNA的表达较健康组增高(P＜0.01).APS可呈剂量依赖性减少LPS诱导的TLR4 mRNA高表达,抑制LPS诱导的TLR4蛋白高表达和IκBα蛋白的降解(P＜0.05,P＜0.01,P＜0.001).结论 TLR-NF-κB信号途径介导的炎症反应和免疫紊乱是高血压病血管内皮损伤的机制之一,APS可通过抑制其表达保护血管内皮细胞损伤.     

长链非编码RNA RP11-490M8.1对脂多糖诱导的人脐静脉内皮细胞炎症的影响

目的 探讨长链非编码RNA(LncRNA)RP11-490M8.1在内皮细胞中对炎症因子的调控作用及其对动脉粥样硬化发生、发展的影响。方法 收集3对AS斑块组织和动脉内膜正常组织进行基因芯片分析,挖掘表达显著异常的基因;用脂多糖(lipopolysaccharide, LPS)浓度梯度和时间梯度处理内皮细胞、构建慢病毒载体过表达LncRNA RP11-490M8.1,以实时荧光定量PCR(qRT-PCR)检测LncRNA RP11-490M8.1和细胞炎症因子如肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、核转录因子-κB(NF-κB)的RNA水平,以免疫印迹(western blot)法检测细胞炎症因子(IL-6、TNF-α、NF-κB)的蛋白表达水平。最后将内皮细胞接种于6孔细胞培养板中,分为4组进行干预：(1)慢病毒空载体对照组(对照组):慢病毒空载体(LV-MOCK)转染内皮细胞培养24 h;(2)慢病毒空载体加LPS(LV-MOCK+LPS):LV-MOCK转染内皮细胞培养6 h,加1 000 ng/mL的LPS培养18 h;(3)慢病毒过表达组(LV-LncR...     

炎症反应及细胞内钙浓度异常升高在脓毒症发展中的作用分析

探讨炎症反应及细胞内钙浓度异常升高(钙超载)在脓毒症发展中的作用。构建血管内皮细胞损伤模型,后使用基质相互作用分子1(Stim1)抑制表达载体、Tlr4抑制表达载体转染细胞,再通过脂多糖活化Toll样受体4(Tlr4)抑制表达载体转染12 h,脂多糖刺激6 h,使用核转录因子-κB(NF-κB)抑制剂PDTC 以及Tlr4抑制表达载体转染1 h、12 h。结果显示脂多糖刺激人脐静脉内皮细胞后Tlr4、髓质分化蛋白2、NF-κB mRNA相对表达量明显高于刺激前(P<0.05)。Stim1抑制表达载体转染可显著逆转脂多糖诱导的人脐静脉内皮细胞外钙离子内流。Tlr4抑制表达载体+脂多糖组、Tlr4及Stim1抑制表达载体+脂多糖组钙离子浓度明显低于脂多糖组(P<0.05)。 实验表明,下调Tlr4、NF-κB、Stim1表达可显著抑制脂多糖诱导的血管内皮细胞炎症反应和钙超载,缓解脓毒症病情进展。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.